title: Npc1 is involved in sterol trafficking in the filamentous fungus Fusarium graminearum.
abstract: The ortholog of the human gene NPC1 was identified in the plant pathogenic, filamentous fungus Fusarium graminearum by shared amino acid sequence, protein domain structure and cellular localization of the mature fungal protein. The FusariumNpc1 gene shares 34% amino acid sequence identity and 51% similarity to the human gene, has similar domain structure and is constitutively expressed, although up-regulated in ungerminated macroconidia and ascospores. GFP-tagged Npc1p localizes to the fungal vacuolar membrane. Cultures derived from a Δnpc1 mutant strain contain significantly more ergosterol than cultures of the wildtype. Staining with the fluorescent, sterol binding dye filipin, shows that ergosterol accumulates in vacuoles of the Δnpc1 mutant but not the wildtype strain. The Δnpc1 mutant has a temperature dependent reduction in growth and greater sensitivity to the ergosterol synthesis inhibiting fungicide tebuconazole compared with the wildtype strain or the mutant complemented with wildtype Npc1. The mutant also is significantly reduced in pathogenicity to wheat. Our results are consistent with the interpretation that Npc1p is important for normal transport of ergosterol from the vacuole and is essential for proper membrane function under particular environmental conditions.
authors.affiliation: Department of Plant Pathology, University of Minnesota, St. Paul, MN 55108, USA. Andy.Breakspear@bbsrc.ac.uk
authors.forename: Andrew
authors.initials: A
authors.lastname: Breakspear
authors.forename: Matias
authors.initials: M
authors.lastname: Pasquali
authors.forename: Karen
authors.initials: K
authors.lastname: Broz
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Dong
authors.forename: H Corby
authors.initials: HC
authors.lastname: Kistler
mesh_terms: D019677:Artificial Gene Fusion; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007425:Intracellular Membranes / Q000737:chemistry; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013261:Sterols / Q000378:metabolism*; D014617:Vacuoles / Q000737:chemistry
chemical_list: D005656:Fungal Proteins; D013261:Sterols; D049452:Green Fluorescent Proteins